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in situs: Fixed and store for too long?

Started by plasmodel, Jun 21 2009 04:30 PM

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#1 plasmodel

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Posted 21 June 2009 - 04:30 PM

Hi,

I will be doing in situs soon. Is it ok if I use embryos/larva that have been fixed and were stored in MeOH (4C) several months ago? Will it affect any results?

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#2 cellcounter

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Posted 09 July 2009 - 08:51 AM

plasmodel, on Jun 21 2009, 05:30 PM, said:

Hi,

I will be doing in situs soon. Is it ok if I use embryos/larva that have been fixed and were stored in MeOH (4C) several months ago? Will it affect any results?

Mouse embryos fixed 4% PFA overnight and stored in MeOH at -20'C (without freezer malfuntion or embryo drying) are good for in situ even one year after dissection. Not sure about other spp. 4'C seems risky proposition, but you can try it out. It also depends upon the message level of the gene you are doing the insitus for. If the original message was very high, chances are that you will still detect it.